Cutting Edge: Microbial Products Elicit Formation of Dendritic Cell Aggresome-Like Induced Structures in Macrophages
J Immunol Canadien et al.
174: 2471
Files in this Data Supplement:
Video 1 (QuickTime, 1,778 KB)
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RAW 264.7 macrophages expressing GFP-ubiquitin were treated with 1 μg/mL LPS
from S. typhimurium for 8 h. Coverslips were then mounted onto an inverted Leica DMIRE2
spinning disk confocal microscope with a heated stage and an Orca AG CCD camera.
Approximately 27 confocal images were acquired at different focal planes (0.38 μm optical
slices) through each cell at a given timepoint. Images were acquired every 90 seconds through
the duration of each experiment. All images were obtained with cells kept at 37°C in RPMI
media buffered to pH 7.2 with HEPES. As shown, ubiquitinated protein aggregates move rapidly
in the cytosol and undergo fusion after coming into contact with each other.
Video 2 (QuickTime, 2,185 KB)
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RAW 264.7 macrophages expressing GFP-ubiquitin were treated with 1 μg/mL LPS
from S. typhimurium for 8 h and visualized as in Video 1. Where indicated, nococazole (5 mM)
was added to cells to disrupt microtubules. This treatment rapidly blocked the movement of
DALIS within cells.